Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR): A critical overview on the most promising applications of molecular scissors in oral medicine

The scientific community is continuously working to translate the novel biomedical techniques into effective medical treatments. CRISPR-Cas 9 system (Clustered Regularly Interspaced Short Palindromic Repeats-9), commonly known as the “molecular scissor”, represents a recently developed biotechnology able to improve the quality and the efficacy of traditional treatments, related to several human diseases, such as chronic diseases, neurodegenerative pathologies and, interestingly, oral diseases. Of course, dental medicine has notably increased the use of biotechnologies to ensure modern and conservative approaches: in this landscape, the use of CRISPR-Cas-9 system may speed and personalize the traditional therapies, ensuring a good predictability of clinical results. The aim of this critical overview is to provide evidence on CRISPR efficacy, taking into specific account its applications in oral medicine.     

PRELIMINARY EVALUATION OF THE APPLICATION OF THE FTIR SPECTROSCOPY TO CONTROL THE GEOGRAPHIC ORIGIN AND QUALITY OF VIRGIN OLIVE OILS

ABSTRACT

A rapid Fourier transform infrared (FTIR) attenuated total reflectance spectroscopic method was applied to determine qualitative parameters such as free fatty acid (FFA) content and the peroxide value (POV) in virgin olive oils. Calibration models were constructed using partial least squares regression on a large number of virgin olive oil samples. The best results (R² = 0.955, root mean square error in cross validation [RMSECV] = 0.15) to evaluate FFA content expressed in oleic acid % (w/w) were obtained considering a calibration range from 0.2 to 9.2% of FFA relative to 190 samples. For POV determination, the result obtained, built on 80 olive oil samples with a calibration range from 11.1 to 49.7 meq O₂/kg of oil, was not satisfactory (R² = 0.855, RMSECV = 3.96). We also investigated the capability of FTIR spectroscopy, in combination with multivariate analysis, to distinguish virgin olive oils based on geographic origin. The spectra of 84 monovarietal virgin olive oil samples from eight Italian regions were collected and elaborated by principal component analysis (PCA), considering the fingerprint region. The results were satisfactory and could successfully discriminate the majority of samples coming from the Emilia Romagna, Sardinian and Sicilian regions. Moreover, the explained variance from this PCA was higher than 96%.

PRACTICAL APPLICATIONS

The verification of the declared origin or the determination of the origin of an unidentified virgin olive oil is a challenging problem. In this work, we have studied the applicability of Fourier transform infrared coupled with multivariate statistical analysis to discriminate the geographic origin of virgin olive oil samples from different Italian regions.

     

Long term immunological perturbations post DAA therapy in chronic HCV/HIV co-infected patients

Direct-acting antiviral (DAA) therapies are efficacious for the achievement of sustained virologic response (SVR) in almost all treated hepatitis C virus (HCV)-infected patients. However, the impacts of HCV eradication on immune function and chronic immune activation in the long-term remain controversial and limited, especially in patients co-infected with human immunodeficiency virus (HIV). Indeed, although restoration of many immune responses clearly can be observed, several features of immune perturbations persist over time after HCV clearance. Understanding the degree and reasons of the partial recovery of the immune system in chronic HCV/HIV co-infection after HCV elimination is pivotal to avoid disease progression and possible long-term clinical outcomes in cured patients, as well as contributing to the development of immunotherapy drug design.     

Mesenchymal stem cell secretome and nanotechnology: Combining therapeutic strategies

Mesenchymal stem cells (MSC) have pushed the field of stem cell-based therapies by inducing tissue regeneration, immunosuppression, and angiogenesis mainly through vesicles and soluble factors release (paracrine signaling). MSC-extracellular vesicles (MSC-EV) adaptable secretome and homing to injured sites allowed researchers to unlock a new era of cell-free based therapy. In parallel, nanoparticles (NP) have been explored in contributing to transport and drug delivery systems, giving drugs desired physical-chemical properties to exploit cell behavior. However, NPs can be quickly recognized by immune cells and cleared from circulation. In this viewpoint, we explore how combining both therapeutic strategies can improve efficacy and circumvent limitations of both therapies. MSCEV benefit from the potent MSC membrane composition, guiding chemotaxis to tumor sites, a very restricted microenvironment. MSC-EV has low immunogenicity, high stability, long half-life and can explore tissue targeting ligands as a precise drug carry, even across biological barriers. Those properties promote enhanced targeted drug delivery that can be combined with NP, exploring biological membrane production through: 1. direct cell therapy with NP-infused MSC; 2. NP-containing MSC-EV generated by NP-infused MSC; 3. by coating NP in MSC membrane (“MSC NanoGhosts”), allowing precise cargo definition without losing targeting. Therefore, nanotechnology combined with cell-based therapeutic resources can greatly improve targeted drug delivery, improving efficacy and opening a new venue of therapeutic possibilities.     

DIFFERENTIAL SCANNING CALORIMETRY DETECTION OF HIGH OLEIC SUNFLOWER OIL AS AN ADULTERANT IN EXTRA-VIRGIN OLIVE OIL

Differential scanning calorimetry (DSC) was employed to detect high oleic sunflower oil (HOSo) as an adulterant in extra-virgin olive oil (EVOo) by means of cooling and heating thermograms. Addition of HOSo did not significantly alter cooling profiles of EVOo except for onset temperature of crystallization, which was significantly shifted toward lower temperature at 40% of adulterant addition. At the same percentage of adulteration, the heating profile of EVOo was significantly changed as the major endotherm broadened and the minor event became smaller and less evident. Cooling thermograms of pure oils and their admixtures were deconvoluted into three constituent exothermic peaks in an attempt to detect addition of HOSo at levels lower than 40%. Thermal properties of the two lower-temperature exotherms (area percentage, offset temperature and range of transition) were significantly changed at ≥ 20% of HOSo substitution, suggesting that DSC can be employed to detect this oil as an EVOo adulterant.

PRACTICAL APPLICATIONS

Adulteration of extra-virgin olive oil (EVOo) with cheaper oils from other vegetable sources or seeds, as well as with lower quality olive oils, is a serious concern for oil suppliers and consumers and requires the use of new analytical techniques for their detection. Differential scanning calorimetry (DSC) exhibits some advantages over the classical analytical methods as it does not require sample preparation and use of solvents, thus resulting in a reduced environmental impact. Results suggested that its application to the detection of EVOo adulteration with high oleic sunflower oil, a vegetable oil largely employed for this type of fraud, is promising with the support of the deconvolution analysis of cooling thermograms.